在单细胞水平上实时评估线粒体 DNA 异构动态。

IF 9.4 1区 生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY EMBO Journal Pub Date : 2024-08-05 DOI:10.1038/s44318-024-00183-5
Rodaria Roussou, Dirk Metzler, Francesco Padovani, Felix Thoma, Rebecca Schwarz, Boris Shraiman, Kurt M Schmoller, Christof Osman
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引用次数: 0

摘要

线粒体 DNA(mtDNA)在细胞内有多个拷贝,是线粒体 ATP 生成所必需的。即使在单个细胞内,mtDNA拷贝的序列也可能不同,这种状态被称为异质性。由于无法对这一现象进行实时监测,人们对异质性程度动态变化的基本原理仍然不甚了解。在这里,我们采用基于 mtDNA 的荧光标记、微流控技术和自动细胞追踪技术,在单细胞水平上追踪活的异质酵母群体中的 mtDNA 变异。这种方法与直接 mtDNA 跟踪和数据驱动的数学建模相结合,揭示了细胞分裂过程中 mtDNA 拷贝的非对称分割,以及线粒体融合和分裂频率的限制,是 mtDNA 变异分离的关键驱动力。鉴于我们的方法还有助于评估完整 mtDNA 和突变 mtDNA 之间的分离,我们预计它将有助于阐明 mtDNA 纯化选择的内在机制。
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Real-time assessment of mitochondrial DNA heteroplasmy dynamics at the single-cell level.

Mitochondrial DNA (mtDNA) is present in multiple copies within cells and is required for mitochondrial ATP generation. Even within individual cells, mtDNA copies can differ in their sequence, a state known as heteroplasmy. The principles underlying dynamic changes in the degree of heteroplasmy remain incompletely understood, due to the inability to monitor this phenomenon in real time. Here, we employ mtDNA-based fluorescent markers, microfluidics, and automated cell tracking, to follow mtDNA variants in live heteroplasmic yeast populations at the single-cell level. This approach, in combination with direct mtDNA tracking and data-driven mathematical modeling reveals asymmetric partitioning of mtDNA copies during cell division, as well as limited mitochondrial fusion and fission frequencies, as critical driving forces for mtDNA variant segregation. Given that our approach also facilitates assessment of segregation between intact and mutant mtDNA, we anticipate that it will be instrumental in elucidating the mechanisms underlying the purifying selection of mtDNA.

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来源期刊
EMBO Journal
EMBO Journal 生物-生化与分子生物学
CiteScore
18.90
自引率
0.90%
发文量
246
审稿时长
1.5 months
期刊介绍: The EMBO Journal has stood as EMBO's flagship publication since its inception in 1982. Renowned for its international reputation in quality and originality, the journal spans all facets of molecular biology. It serves as a platform for papers elucidating original research of broad general interest in molecular and cell biology, with a distinct focus on molecular mechanisms and physiological relevance. With a commitment to promoting articles reporting novel findings of broad biological significance, The EMBO Journal stands as a key contributor to advancing the field of molecular biology.
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